mPFC-QuEChERS净化联合UPLC-MS/MS法快速测定3种根茎类中药材中16种真菌毒素

Determination of 16 Mycotoxins in Three Radix and Rhizome Chinese Herbal Medicines by mPFC-QuEChERS with UPLC-MS/MS

建立了快速滤过型净化柱(mPFC-QuEChERS)结合超高效液相色谱-串联质谱(UPLC-MS/MS)同时测定黄芪等3种根茎类中药材中16种真菌毒素的分析方法。通过优化色谱和质谱条件及样品前处理方法,确定样品经80%乙腈(含2%乙酸)高速匀浆提取,快速滤过型净化柱单步净化,shim-pack Velox PFPP色谱柱分离,以甲醇和0.01%甲酸/0.2 mmol/L甲酸铵水溶液为流动相进行梯度洗脱。采用电喷雾(ESI)离子源,在多反应监测(MRM)模式下以正负模式同时扫描采集,基质匹配标准曲线外标法进行定量。结果表明,16种真菌毒素在一定的质量浓度范围内线性关系良好(r>0.99),方法检出限(LOD)为0.3~25μg/kg,定量下限(LOQ)为0.5~50μg/kg。在2.5、10、20倍LOQ加标水平下,16种真菌毒素的平均回收率为63.3%~110%,相对标准偏差(RSD)为0.20%~6.7%。对60批中药材中16种真菌毒素进行检测,检出率为30%,检出赭曲霉毒素A(OTA)、15-乙酰基脱氧雪腐镰刀烯醇(15-ACE)和杂色曲霉毒素(ST)。其中9批黄芪、2批党参样品检出OTA,检出率最高。该法操作简便、灵敏度高、实用性强,可用于黄芪等3种根茎类中药材中16种真菌毒素的快速筛查和定量分析。

A method was established using a rapid multiple filtrate purification column(mPFCQuEChERS)combined with ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)for the simultaneous determination of sixteen mycotoxins in three radix and rhizome Chinese herbal medicines.Through optimization of the liquid chromatography conditions,mass spec⁃trometry conditions,and sample pretreatment process,the sample was extracted with a high-speed homogenizer using an 80%acetonitrile solution(containing 2%acetic acid),and single-step purified by a mPFC-QuEChERS purification column.The analytes were separated by a shim-pack Velox PF⁃PP chromatographic column,and eluted with a gradient of 0.01%formic acid/0.2 mmol/L formic ac⁃id ammonium solution and methanol as the mobile phase,and determined in positive and negative electrospray ionization mode in multiple reaction monitoring(MRM)mode.Quantitative analysis was performed using the external standard method.Under the optimal experimental conditions,sixteen mycotoxins showed good linear relations in their mass concentrations(r>0.99),with the detection limits(LODs)ranging from 0.3 to 25μg/kg and quantification limits(LOQs)ranging from 0.5 to 50μg/kg.The residual levels of 16 mycotoxins were spiked at 2.5,10 and 20 times of LOQ with aver⁃age recoveries ranging from 63.3%to 110%and relative standard deviations(RSDs)ranging from 0.20%to 6.7%.60 batches of Chinese herbal medicines were detected,and the detection rate was 30%.Ochratoxin A(OTA),15-acetyldeoxynivalenol(15-ACE)and sterigmatocystin(ST)were relatively high detected.Notably,OTA was detected in 9 batches of Astragali Radix and 2 batches of Co⁃donopsis Radix,representing the highest detection rate.The method was simple,high sensitive,and practicable,which can be used for the rapid screening and quantitative analysis of 16 mycotoxins in radix and rhizome Chinese herbal medicines.